Abstract: In various embodiments, method and devices for delivering large cargos (e.g., organelles, chromosomes, bacteria, and the like) into cells are provided. In certain embodiments method of delivering a large cargo into eukaryotic cells, are provided that involve providing eukaryotic cells disposed on one side of a porous membrane; providing the cargo to be delivered in a solution disposed in a reservoir chamber on the opposite side of the porous membrane; and applying pressure to the reservoir chamber sufficient to pass the cargo through pores comprising said porous membrane wherein said cargo passes through cell membranes and into the cells.

Abstract: An illumination unit includes one or more light sources each including a solid-state light-emitting device having a light emission region configured of one or more light-emission spots, one or more traveling-direction angle conversion device each converting a traveling-direction-angle of light, and an integrator including a first fly-eye lens having cells which receive light from the traveling-direction angle conversion device and a second fly-eye lens having cells which receive light from the first fly-eye lens, the integrator uniformalizing illumination distribution in a predetermined illumination area. An optical system configured with the traveling-direction angle conversion device and the first and second fly-eye lenses has an optical magnification which allows each of light source images to have a size not exceeding a size of the cell in the second fly-eye lens, the light source images being formed on the second fly-eye lens by the respective cells in the first fly-eye lens.

Abstract: The invention relates to a laser assembly (100) having a laser (L) for generating primary laser pulses (1), beam splitting optics (15) for splitting a primary laser pulse into a plurality of temporally staggered sub-pulses, and having focusing optics (17-19) for focusing the sub-pulses in or on an object (20) so that every sub-pulse is focused in a separate focus volume (F). The invention is characterized in that the mutual spatial and/or temporal relationship of the focus volumes (F) of the sub-pulses originating from a common primary laser pulse is variably adjustable. The invention also relates to a corresponding method.

Abstract: A system and method are described for electroporating a sample that utilizes one or more sets of electrodes that are spaced apart in order to hold a surface tension constrained sample between the electrodes. The first electrode is connected to the lower body of the system while the second electrode is connected to the upper body. Both electrodes are connected to a pulse generator. Each electrode has a sample contact surface such that the first electrode and the second electrode may be positioned to hold a surface tension constrained sample between the two sample contact surfaces and the sample may receive a selected electric pulse.

Abstract: Method for the selection or the processing of first particles sensitive to the application of an external stimulus including the step of producing, through the application of the external stimulus, the permeabilization of at least a selected first particle, consisting in the organization of the first particles through a first force field, to generate a second force field substantially placed in proximity of at least a selected first particle to be permeabilized.

Abstract: A device for use in laser optical transfection of biological targets including an optofluidic microdevice and a piece of optical glass. The optofluidic microdevice has a central vertical outlet and a microchannel network that includes a plurality of entrapping channels with narrowings. The microchannel network is fused with the optical glass. In one aspect the device is used with a petri dish having an optical window. In another aspect the device is used with a well plate having a plurality of wells and associated optical windows. In a third aspect the device is used with a barrier. Each of the aspects forms a peripheral space around the optofluidic microdevice capable of retaining a live culture of biological targets and material that is desired to be injected into those biological targets. Polymer tubing is inserted into the central vertical outlet which connects the device to an external pump.

Abstract: The invention, in some aspects relates to compositions and methods for altering cell activity and function and the introduction and use of light-activated ion channels.

Abstract: Embodiments of the present disclosure provide a multistage procedure for treatment of biological samples (e.g., living cells with membranes, and the like) with a substance (e.g., a drug, DNA, RNA, plasmids, and other biomolecules or materials) to achieve more efficacious intracellular delivery and transfection.

Abstract: The present invention includes methods for effecting phenotype conversion in a cell by transfecting the cell with phenotype-converting nucleic acid. Expression of the nucleic acids results in a phenotype conversion in the transfected cell. Preferably the phenotype-converting nucleic acid is a transcriptome, and more preferably an mRNA transcriptome.

Abstract: A method of delivering exogenous molecules, comprising: providing a plurality of cells having a cell membrane; adding a plurality of exogenous molecules to the cells; exposing the cells to a defocused infrared (IR) light to permeabilize the cell membrane of the cells; and delivering the exogenous molecules to the cells through the permeablized cell membrane, wherein an intensity of the IR light at the optical focus is at least greater than or equal to an order of 104 W/cm2.

Abstract: The present disclosure is directed to novel polynucleotide sequences for use in Nannochloropsis gaditana. The novel polynucleotide sequences include control sequences and coding sequences. Also disclosed are novel gene expression constructs wherein N. gaditana promoters/control regions are operatively linked to N. gaditana or non-N. gaditana coding sequences. These novel polynucleotide sequences and expression constructs can be introduced into N. gaditana and can recombine into the N. gaditana genome. Expression from these polynucleotide sequences and expression constructs can enhance N. gaditana biomass and/or lipid biosynthesis. Also disclosed are methods for modifying N. gaditana, for example by stably transforming N. gaditana with nucleic acid sequences, growing the modified N. gaditana, and obtaining biomass and biofuels from the modified N. gaditana.

Abstract: The invention relates to an apparatus for introducing a biological material, a method of introducing a biological material, and a magnetic support for introducing a biological material with the object of providing an apparatus for introducing a biological material, a method of introducing a biological material, and a magnetic support for introducing a biological material whereby a biological material can be efficiently introduced into a host. The invention comprises: one or more packing units in which a mixture solution containing a large number of magnetic supports carrying a biological material to be introduced into a host such as cells upon using, together with a large number of the hosts in a liquid is pooled; and an introduction treatment unit in which a magnetic force affecting the inside of the packing unit is controlled so as to move the magnetic supports relatively with respect to the host so that the biological material can be introduced into the host.

Abstract: The present invention relates to a cell fusion chamber in which two types of cells having different diameters are fused, the cell fusion chamber including: a cell fusion region in which cell fusion is carried out; a pair of electrodes formed by a conductor and disposed opposite to each other in the cell fusion region; and a partition wall having at least one fine pore; near the fine pore, a cell fusion device including a cell fusion container containing a cell fusion region; a pair of electrodes; a spacer; and an insulator disposed between the spacer and one of the electrodes and having at least one fine pore; and an electronic power supply which applies an alternating voltage and a voltage pulsed direct current to the electrodes, and a cell fusion method using the same.

Abstract: The present invention provides a sonoporation-based method that can be universally applied for delivery of compounds into Gram positive bacteria. Gram positive bacteria which can be transformed by sonoporation include, for example, Bacillus, Streptococcus, Acetobacterium, and Clostridium. Compounds which can be delivered into Gram positive bacteria via sonoporation include nucleic acids (DNA or RNA), proteins, lipids, carbohydrates, viruses, small organic and inorganic molecules, and nano-particles.

Abstract: The invention relates to an apparatus for introducing a biological material, a method of introducing a biological material, and a magnetic support for introducing a biological material with the object of providing an apparatus for introducing a biological material, a method of introducing a biological material, and a magnetic support for introducing a biological material whereby a biological material can be efficiently introduced into a host. The invention comprises: one or more packing units in which a mixture solution containing a large number of magnetic supports carrying a biological material to be introduced into a host such as cells upon using, together with a large number of the hosts in a liquid is pooled; and an introduction treatment unit in which a magnetic force affecting the inside of the packing unit is controlled so as to move the magnetic supports relatively with respect to the host so that the biological material can be introduced into the host.

Abstract: Hemoglobin in a sample solution is quickly and reliably denatured; at the same time, quick and accurate measurement of hemoglobin and a hemoglobin derivative is realized. In a method for measuring hemoglobin and a hemoglobin derivative, and a reagent composition, a measurement kit, an analysis device, and an analysis system used in the method, a sample solution containing a blood component is treated with a nonionic surfactant, an oxidizing agent, and a metal salt to denature hemoglobin in the sample solution to measure the hemoglobin, and thereafter the amount of a hemoglobin derivative in the sample is measured by an immunological method using an antibody specifically binding to a denatured site of the denatured hemoglobin derivative.

Abstract: A method for identifying a molecule that binds an irradiated tumor in a subject and molecules identified thereby. In some embodiments, the method includes the steps of (a) exposing a tumor to ionizing radiation; (b) administering to a subject a library of diverse molecules; and (c) isolating from the tumor one or more molecules of the library of diverse molecules, whereby a molecule that binds an irradiated tumor is identified. Also provided are targeting ligands that bind an irradiated tumor and therapeutic and diagnostic methods that employ the disclosed targeting ligands.

Abstract: Methods and systems for acoustically treating material using a continuous process in which material may be caused to flow in a continuous or intermittent fashion into/out of an acoustic treatment chamber where the material is exposed to focused acoustic energy. The methods and systems may be arranged to permit continuous processing for extended periods while an acoustic energy source operates at a relatively high power output. Treatment chambers may include features such as an acoustic window, a heat exchanger, inlet/outlet flow arrangements, an inspection window, insert elements that define a treatment volume size or shape, etc. Treatment system configurations relating to arrangements of a treatment chamber relative to an acoustic source and coupling medium, material flow paths, and others are provided.

Abstract: The invention relates to a method for subjecting adherent cells to at least one electric field, in which the electric field is generated by applying a voltage to at least two active electrodes 63, wherein at least three electrodes 63, 64 are provided, and wherein at least two electrodes 63, 64 are active electrodes 63 when the voltage is applied in order to generate a first electric field, and in which at least one second electric field is generated, wherein at least one of the two previously active electrodes 63 is a potential-free electrode 64 when the voltage is applied.

Abstract: Systems and methods for magnetic targeting of therapeutic particles are provided. Therapeutic particles comprise one or more magnetic or magnetizable materials and at least one therapeutic agent. Therapeutic particles are specifically targeted using uniform magnetic fields capable of magnetizing magnetizable materials, and can be targeted to particular locations in the body, or can be targeted for capture, containment, and removal. Therapeutic particles can comprise antioxidant enzymes, and can be targeted to cells to protect the cells from oxidative damage.

Abstract: A method for facilitating a delivery of a molecule into an interior space of a cell includes the steps of introducing a molecule into a biological structure comprising a cell and applying a substantially continuous low-level electric field, in the form of non-thermal plasma (ionized gas) generated by a direct current voltage applied to an electrode, to the molecule and biological structure. The field is applied for a duration sufficient to effect a change in porosity the cell of the biological structure sufficient to facilitate an entry of a desired molecule into an interior thereof.

Abstract: A method of applying a nondestructive mechanical force on one or more cells in aqueous environment by inducing heat generated acoustic pressure pulses. The method comprises providing an energy transmission pattern to induce the applying of a desired nondestructive mechanical force on a at least one cell by forming a plurality of heat generated acoustic pressure pulses in an aqueous environment and instructing the radiation of a target area in proximity to the at least one cell in the aqueous environment with light energy according to the energy transmission pattern.

Abstract: A microfluidic cartridge for isolating biological molecules having a capture chamber containing functionalized solid supports maintained in a fluidized state provides reduced pressure drops and bubble formation during microfluidic extraction. The cartridge may include an electric field lysis chamber and/or a chemical lysis chamber. The electric-field lysis chamber may comprise an electrically insulating structure arranged between two opposing planar electrodes.

Abstract: Disclosed is a liquid culture medium for substance introduction, which is capable of increasing the survival rate of cells after substance introduction as much as possible when the cells are irradiated with plasma for the purpose of introducing a target substance into each of the cells. Specifically disclosed is a liquid culture medium for substance introduction, which is used for the purpose of introducing a predetermined target substance into a cell and enables introduction of the target substance into the cell by having the cell in the liquid culture medium, which contains the target substance, irradiated with a plasma jet. The liquid culture medium contains a damage preventing component that prevents the cell from damage due to the plasma jet.

Abstract: A method of delivering exogenous molecules, comprising: providing a plurality of cells having a cell membrane; adding a plurality of exogenous molecules to the cells; exposing the cells to a defocused infrared (IR) light to permeabilize the cell membrane of the cells; and delivering the exogenous molecules to the cells through the permeablized cell membrane, wherein an intensity of the IR light at the optical focus is at least greater than or equal to an order of 104 W/cm2.

Abstract: A method of optoperforation of the membrane of a cell by application of laser pulses characterized by focusing the pulsed laser beam onto the cell membrane to be perforated, applying a series of laser pulses of predetermined pulse energy, measuring the oscillation time of the bubbles formed in the laser focus from the change in laser intensity of a test laser beam transmitted through the laser focus and caused by the bubbles in the laser focus, and increasing the pulse energy to a level at which the oscillation time of the bubbles attains a predetermined value.

Abstract: Disclosed is a method for parallel delivery of agents to and/or into a cell structure, wherein at least two electrolyte-filled tubes are provided together with a counter electrode, the tubes being connected to a voltage or current generator, said agents being introduced into the electrolyte solution contained in the tubes, which are placed close to the cell structure, whereupon the agents are transported through the tubes to said cell structure and into the said structure through pores which have been formed by application of an electric field focused on the cell structure, resulting in electroporation of the cell structure. Also different applications of the method is disclosed, e.g. use of the method in order to transfer cell-impermeant solutes, such as drugs or genes, into the cell structure or out of the cell structure.

Abstract: In method of injecting a substance into a living cell having a cell membrane, the substance, the cell and a liquid are placed into a tapering passage. Energy is applied to the cell, thereby inducing poration. To sort cells, a cellular suspension is placed in a tapering passage, including a narrow end that defines an opening that has a dimension corresponding to a cell size. An acoustic wave is applied, thereby forcing cells having a cell size smaller than the selected cell size through the opening, with a portion of the cells having a cell size not smaller than the selected cell size not forced through the opening. To extract material from a cell, an electric field and an acoustic wave are applied, thereby causing the cell membrane to allow the material to pass out of the cell.

Abstract: A micromachined patch-clamp apparatus is disclosed for holding one or more cells and providing electrical, chemical, or mechanical stimulation to the cells during analysis with the patch-clamp technique for studying ion channels in cell membranes. The apparatus formed on a silicon substrate utilizes a lower chamber formed from silicon nitride using surface micromachining and an upper chamber formed from a molded polymer material. An opening in a common wall between the chambers is used to trap and hold a cell for analysis using the patch-clamp technique with sensing electrodes on each side of the cell. Some embodiments of the present invention utilize one or more electrostatic actuators formed on the substrate to provide mechanical stimulation to the cell being analyzed, or to provide information about mechanical movement of the cell in response to electrical or chemical stimulation.

Abstract: A tissue sample preservation process and device for improving and standardizing tissue preservation procedure, including placing tissue specimens in a cold fixative, performing fixative penetration at a refrigerated temperature, and accelerating fixative penetration by ultrasound. Also disclosed is the application of ultrasound and temperature control in the dehydration, clearing, and impregnation steps.

Abstract: The present application includes systems and methods for identifying a compound capable of interacting with a G-Protein Coupled Receptor (GPCR) or Receptor Tyrosine Kinase (RTK) including providing a device capable of measuring cell-substrate impedance operably connected to an impedance analyzer, adding test cells expressing a GPCR or a RTK to wells of the device, measuring first impedances of the wells and optionally determining first cell indices from the first impedances, adding a compound to at least one well containing test cells to form at least one compound well and adding a vehicle control to at least another well containing test cells to form at least one control well, measuring second impedances of the compound well and the control well and optionally determining second cell indices from the second impedances, determining the change in the impedance or cell index for the compound well and the one control well, comparing the change in impedance or cell index between the compound well and the control

Abstract: An extracellular potential measuring device includes a plate portion having a first surface and a second surface opposite to the first surface, and an electrode provided on the second surface of the plate portion. In the plate portion, a pocket having an opening which opens to the first surface is formed, and a through-hole communicating to the second surface from the pocket. The through-hole communicates from a position which is closer to the opening than a deepest point of the first pocket. The electrode is provided around of the opening of the through-hole. In this device, even if a cell to be examined does not reach the deepest point of the pocket, a cell membrane of the cell can tightly attaches onto the through-hole securely without a clearance. Hence, culture solution inside the through-hole is isolated from culture solution over an upper surface of the plate portion, thereby allowing electrochemical changes caused by activities of the cell to be detected efficiently with a detector electrode.

Abstract: The invention relates to a method of viral inactivation by dry heating of a virus present or potentially present in a biological product that has been dried according to the glass transition temperature.

Abstract: A device for measuring an extracellular potential of a test cell includes a substrate having a well formed in a first surface thereof and a first trap hole formed therein. The well has a bottom. The first trap hole includes a first opening formed in the bottom of the well and extending toward a second face of the substrate, a first hollow section communicating with the first opening via a first connecting portion, and a second opening extending reaching the second surface and communicating with the first hollow section via a second connecting portion. The first connecting portion has a diameter smaller than a maximum diameter of the first hollow section, greater than a diameter of the second connecting portion, and smaller than a diameter of the test cell. The device can retain the test cell securely and accept chemicals and the test cell to be put into the device easily.

Abstract: The present invention relates to methods of measuring electrical properties of a cell using electrode devices comprising tapered nanotips having submicrometer dimensions (“nanoelectrodes”) for insertion into a cell. The devices are used to measure electrical properties of the cell and, optionally, may be used to electroporate, the cell or subcellular structures within the cell. The invention also provides arrays of electrode devices having nanotips for simultaneously or sequentially measuring the electrical properties of cells (e.g., such as surface immobilized cells). The electrodes can be used to measure properties of ion channels and in HTS assays to identify drugs which affect the properties of ion channels. The invention additionally provides microfluidic systems adapted for use with the electrode devices having nanotips. In combination with the electrodes, the microfluidic systems provide cell-based biosensors for monitoring cellular responses to conditions, such as exposure to candidate drugs.

Abstract: Method and apparatus for processing a cell culture are provided. The method includes establishing a cell culture within a holding device having one or more wells, each well holding a cell culture, and including a well substrate with at least one electrode in contact with the cell culture; periodically applying at least one electrical pulse to the at least one electrode to prevent cells from attaching to and achieving confluence over the at least one electrode while allowing cells to attach to and achieve confluence over other portions of the well substrate; and discontinuing the periodically applying of the at least one electrical pulse to the at least one electrode after cells have achieved confluence over the other portions of the well substrate, and thereafter, monitoring the cell culture to monitor migration of cells over the electrode(s) from the other portions of the well substrate.

Abstract: A method of treating biocells includes the steps of: a. providing biocells; b. applying at least one stressor to the biocells sufficient to cause nonlethal and reparable cell wall damage to the biocells, thereby putting the biocells in a catabolic state during which catabolic metabolic functions predominate over anabolic metabolic functions; and c. obtaining at least one product produced by the biocells during the catabolic state. In another embodiment, the method includes the steps of: a. providing biocells that are mammalian cells; b. applying at least one stressor to the biocells sufficient to cause nonlethal and reparable cell wall damage to the biocells, the reparable cell wall damage comprising openings that allow increased passage of materials through the cells walls; and c. inserting foreign DNA through the openings into the biocells.

Abstract: The invention is directed to a method for the manipulation of at least one cell, the method comprising the steps of depositing a metal onto the surface of a substrate, placing the at least one cell at or near the surface of the substrate, and irradiating the surface of the substrate with at least one laser pulse. The inventive method is characterized by the formation of surface structures with a size of one micrometer or less on the surface of the substrate prior to depositing the metal thereon. The invention is also directed to a system for the manipulation of at least one cell, the system comprising a substrate with surface structures having a size of 1 micrometer or less, wherein a metal is deposited on the surface structures, and wherein the system further comprises a laser for irradiating the surface structures.

Abstract: A method for introducing biologically active molecules into animal or human cells using an electric current includes suspending the cells and dissolving the biologically active molecules in a buffer solution including HEPES and at least 10 mmol×1?1 magnesium ions (Mg2+), the buffer solution having a buffer capacity of at least 20 mmol×1?1 ×pH?1 at a change in the pH from pH 7 to pH 8 and at a temperature of 25° C., and an ionic strength of at least 200 mmol×1?1. An electric voltage is applied to the suspension.

Abstract: The present invention includes methods for transferring a multigenic phenotype to a cell by transfecting, preferably by phototransfection, and locally transfecting a cell or a cellular process with a laser while the cell is bathed in a fluid medium comprising two or more nucleic acids, thereby introducing the nucleic acid into the interior of the cell. Expression of the nucleic acids results in a multigenic phenotype in the tranfected cell.

Abstract: The invention relates to a novel circuit arrangement for electrotransfection or electrofusion, which enables the transportation of DNA and/or other biologically active molecules to the nucleus of higher eukaryotic cells or the fusion of cells, independent of cell division and with reduced cell mortality.

Abstract: Methods, tip assemblies and kits are provided for introducing material into cells. The tip assemblies include an attachment portion, a channel portion, and a constriction that function to reduce fluid pressure as a fluid passes through the constriction portion from the channel portion, whereby the tip assemblies form pores in the membranes of cells and introduce material into the cells. The material includes for example one selected from the group of: an inorganic compound, a drug, a genetic material, a protein, a carbohydrate, a synthetic polymer, and a pharmaceutical composition.

Abstract: A ready-to-use electroporation cuvette is provided that includes a cuvette, first and second electrodes positioned within the cuvette and electroporation competent cells frozen in a suspension solution within the cuvette, wherein the electroporation cuvette is configured to permit electroporation of the cells when the cells are thawed. The electroporation cuvette may be sealed with a cap that may be color coded to aid the user.

Abstract: The invention concerns a container 1 with chambers 2 which each comprise at least one pair of electrodes including a first 4 and a second electrode 5 for the application of electric voltage for generating an electric field within one chamber 2. At least two first electrodes 4 of different chambers 3 are conductively coupled and at least one second electrode 5 of said chamber 2 is separately conductively connectable. The invention further concerns a method for manufacturing said container 1 as well as a device for electrically contacting at least one of said containers 1.

Abstract: An opening is formed in a cell using laser radiation. A Bessel beam is formed using the laser radiation and the Bessel beam is directed onto the cell to form an opening. The guiding of material towards the opening may be involved using optical trapping/manipulation. The material may change cellular function or analyse cell behaviour. Both pulsed laser radiation and continuous wave radiation may be formed using the same laser.

Abstract: Drug candidate screening methods are applied to discover compounds with activity against ion channel targets. The method may include modulating the transmembrane potential of host cells in a plurality of sample wells with a repetitive application of electric fields so as to set the transmembrane potential to a level corresponding to a pre-selected voltage dependent state of a target ion channel.

Abstract: The introduction of genetic material or molecules of biological interest into cells is a procedure with an increasing interest both for experimental and application purposes, so that electroporation is a widely used technique, but the electroporation of single adhering cells is still impaired. The present application describes an apparatus for the electroporation of any kind of cell adhering to a substrate at any stage of development, where an electrical signal can be driven and applied to a single adhering cell in culture in order to obtain its electroporation. The method to electroporate a single adhering cell with the apparatus of the invention is also described.

Abstract: The present invention provides a method for introducing a molecule into the cytosol of a cell in which the cell is contacted with a photosensitising agent, the cell is irradiated with light of a wavelength effective to activate the photosensitising agent and, substantially at the same time or after the irradiation, the cell is contacted with the molecule to be introduced, particularly for use in cancer treatment, gene therapy and vaccination.

Abstract: A method for introducing biologically active molecules into animal or human cells using electric current includes suspending the cells and dissolving the biologically active molecules in a buffer solution which has a buffer capacity of at least 20 mmol×l?1×pH?1 and an ionic strength of at least 200 mmol×l?1 at a change in the pH from pH 7 to pH 8 and at a temperature of 25° C. to form a suspension. The method further includes applying an electric voltage to the suspension so as to introduce the biologically active molecules into animal or human cells.

Abstract: Optoinjection method for transiently permeabilizing a target cell by (a) illuminating a population of cells contained in a frame; (b) detecting at least one property of light directed from the frame; (c) locating a target cell by the property of light; and (d) irradiating the target cell with a pulse of radiation.